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旋转壁式生物反应器动态条件下支架微观和宏观结构对雪旺细胞增殖的影响

Impact of Scaffold Micro and Macro Architecture on Schwann Cell Proliferation under Dynamic Conditions in a Rotating Wall Vessel Bioreactor.

作者信息

Valmikinathan Chandra M, Hoffman John, Yu Xiaojun

机构信息

Department of Chemistry, Chemical Biology and Biomedical Engineering Stevens Institute of Technology, Hoboken, NJ, 07030.

出版信息

Mater Sci Eng C Mater Biol Appl. 2011 Jan 1;31(1):22-29. doi: 10.1016/j.msec.2010.04.001.

Abstract

Over the last decade tissue engineering has emerged as a powerful alternative to regenerate lost tissues owing to trauma or tumor. Evidence shows that Schwann cell containing scaffolds have improved performance in vivo as compared to scaffolds that depend on cellularization post implantation. However, owing to limited supply of cells from the patients themselves, several approaches have been taken to enhance cell proliferation rates to produce complete and uniform cellularization of scaffolds. The most common approach is the application of a bioreactor to enhance cell proliferation rate and therefore reduce the time needed to obtain sufficiently significant number of glial cells, prior to implantation.In this study, we show the application of a rotating wall bioreactor system for studying Schwann cell proliferation on nanofibrous spiral shaped scaffolds, prepared by solvent casting and salt leaching techniques. The scaffolds were fabricated from polycaprolactone (PCL), which has ideal mechanical properties and upon degradation does not produce acidic byproducts. The spiral scaffolds were coated with aligned or random nanofibers, produced by electrospinning, to provide a substrate that mimics the native extracellular matrix and the essential contact guidance cues.At the 4 day time point, an enhanced rate of cell proliferation was observed on the open structured nanofibrous spiral scaffolds in a rotating wall bioreactor, as compared to static culture conditions. However, the cell proliferation rate on the other contemporary scaffolds architectures such as the tubular and cylindrical scaffolds show reduced cell proliferation in the bioreactor as compared to static conditions, at the same time point. Moreover, the rotating wall bioreactor does not alter the orientation or the phenotype of the Schwann cells on the aligned nanofiber containing scaffolds, wherein, the cells remain aligned along the length of the scaffolds. Therefore, these open structured spiral scaffolds pre-cultured with Schwann cells, in bioreactors could potentially shorten the time needed for grafts for peripheral nerve regeneration.

摘要

在过去十年中,组织工程已成为一种强大的替代方法,用于再生因创伤或肿瘤而受损的组织。有证据表明,与依赖植入后细胞化的支架相比,含有雪旺细胞的支架在体内具有更好的性能。然而,由于患者自身细胞供应有限,人们采取了多种方法来提高细胞增殖率,以实现支架的完全均匀细胞化。最常见的方法是应用生物反应器来提高细胞增殖率,从而减少在植入前获得足够数量神经胶质细胞所需的时间。在本研究中,我们展示了旋转壁生物反应器系统在研究雪旺细胞在通过溶剂浇铸和盐沥滤技术制备的纳米纤维螺旋形支架上增殖方面的应用。这些支架由聚己内酯(PCL)制成,PCL具有理想的机械性能,降解时不会产生酸性副产物。螺旋支架涂覆有通过静电纺丝产生的排列或随机的纳米纤维,以提供一种模仿天然细胞外基质和基本接触导向线索的基质。在第4天时间点,与静态培养条件相比,在旋转壁生物反应器中开放结构的纳米纤维螺旋支架上观察到细胞增殖速率提高。然而,在同一时间点,与静态条件相比,其他当代支架结构(如管状和圆柱形支架)上的细胞增殖速率在生物反应器中显示降低。此外,旋转壁生物反应器不会改变含有排列纳米纤维的支架上雪旺细胞的取向或表型,其中细胞沿支架长度保持排列。因此,在生物反应器中用雪旺细胞预培养的这些开放结构螺旋支架可能会缩短用于周围神经再生移植物所需的时间。

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